Understanding Compartmentalized Leucine Metabolism Downstream of mTORC1 Signaling

了解 mTORC1 信号下游的区室化亮氨酸代谢

• 批准号: 10209938
• 负责人: Paul Rosen
• 金额: $ 4.6万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10209938
• 关键词: Acetyl Coenzyme A; Address; Affect; Amino Acids; Automobile Driving; Biochemical; Biology; Biophysics; Biosensor; Catabolism; Cells; Cellular Metabolic Process; Communication; Complex; Critical Thinking; Cytosol; Development; Diabetes Mellitus; Disease; Engineering; Environment; Enzymes; Epilepsy; Essential Amino Acids; FRAP1 gene; Genetic; Growth; Growth Factor; Human; Institutes; Isotope Labeling; Isotopes; Lead; Leucine; Lysosomes; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Metabolic; Metabolism; Methods; Mitochondria; Nutrient; Oral; Output; Pathology; Pathway interactions; Phosphotransferases; Process; Protein Engineering; Proteins; Proteolysis; Reagent; Regulation; Resources; Role; Signal Transduction; Site; Starvation; Stress; Testing; Time; Training; Work; amino acid metabolism; base; cell growth; design and construction; experience; fluorescence imaging; inhibitor/antagonist; innovation; insight; interdisciplinary approach; interest; metabolomics; novel; novel therapeutic intervention; prototype; response; science education; sensor; skills; tool

Project Summary/Abstract: The mechanistic target of rapamycin complex 1 (mTORC1) kinase coordinates cell growth and metabolism, integrating signals from growth factors, nutrients, and stresses. Many diseases, including cancers, diabetes, and epilepsy, have been associated with aberrations in mTORC1 signaling. Understanding the inputs for and outputs of mTORC1 signaling is of critical importance in driving innovations in new therapeutic interventions for these diseases. Recently, it has been appreciated that mTORC1 signaling is important for compartmentalized amino acid metabolism; this is particularly true with respect to the essential amino acid leucine. While leucine activates mTORC1 in the cytosol, mTORC1 also regulates leucine metabolism in the cytosol, mitochondria, and lysosomes. Recent work from our lab has demonstrated a key role for mTORC1 signaling in regulating the efflux of leucine and other essential amino acids from lysosomes as part of the cellular response to starvation, but how mTORC1 signaling affects leucine levels in the cytosol and mitochondria, the two other important sites of leucine metabolism in cells, remains unknown. Answering this question could help to illuminate how mTORC1 rewires amino acid metabolism in response to starvation. We will determine how mTORC1 signaling affects compartmentalized leucine metabolism through the following specific aims: 1) optimize methods to measure compartmentalized leucine levels, 2) determine how mTORC1 activity changes leucine levels in the cytosol and mitochondria, and 3) characterize how mTORC1 activity affects the rate of leucine catabolism. We will use a multidisciplinary approach, drawing on tools including genetically encoded fluorescent biosensors, organellar metabolomics, and isotope tracing, to systematically address how mTORC1 signaling affects leucine levels in the cytosol and mitochondria. Our results will be important for understanding the role of mTORC1 in regulating adaptive responses to starvation, and may identify previously unappreciated metabolic vulnerabilities in diseases with dysregulated mTORC1 signaling. The proposed work, which will take place at the MIT Biology Department/Whitehead Institute— outstanding intellectual environments that are dedicated to scientific training, includes a strong and comprehensive training plan. This plan emphasizes the development of experimental skills in cutting-edge biochemical approaches including protein engineering, fluorescence imaging, and metabolomics; critical thinking abilities; oral and written scientific communication skills; and science education experience.

项目摘要/摘要： 雷帕霉素复合体1(MTORC1)激酶的机制靶点是协调细胞生长和 新陈代谢，整合来自生长因子、营养物质和压力的信号。许多疾病，包括癌症， 糖尿病和癫痫与mTORC1信号的异常有关。了解输入内容 MTORC1信号的形成和输出对于推动新的治疗方法的创新至关重要 对这些疾病的干预。最近，人们已经意识到mTORC1信号对于 分割氨基酸代谢；对于必需氨基酸来说尤其如此 亮氨酸。当亮氨酸激活胞浆中的mTORC1时，mTORC1也调节细胞内的亮氨酸代谢。 胞浆、线粒体和溶酶体。我们实验室最近的工作证明了mTORC1的关键作用 调节溶酶体内亮氨酸和其他必需氨基酸外流的信号转导 细胞对饥饿的反应，但mTORC1信号如何影响胞浆和细胞内的亮氨酸水平 线粒体是细胞内亮氨酸代谢的另外两个重要部位，目前尚不清楚。回答这个问题 这个问题可能有助于阐明mTORC1是如何改变氨基酸代谢以应对饥饿的。我们 将通过以下方式确定mTORC1信号如何影响间隔化的亮氨酸代谢 具体目标：1)优化测量间隔化亮氨酸水平的方法；2)确定mTORC1 活性改变胞浆和线粒体中的亮氨酸水平，以及3)mTORC1活性如何表征 影响亮氨酸分解代谢的速度。我们将使用多学科方法，利用包括 基因编码的荧光生物传感器、细胞器代谢组学和同位素示踪，以系统地 阐述mTORC1信号如何影响胞浆和线粒体中的亮氨酸水平。我们的结果将是 对于理解mTORC1在调节对饥饿的适应性反应中的作用很重要，并且可能 在mTORC1信号异常的疾病中识别以前未被认识到的代谢脆弱性。 这项拟议的工作将在麻省理工学院生物系/怀特黑德研究所- 致力于科学训练的优秀智力环境，包括强大的 全面的培训计划。该计划强调发展尖端技术的实验技能。 包括蛋白质工程、荧光成像和代谢组学在内的生化方法；关键 思维能力；口头和书面的科学交流能力；以及科学教育经验。